CN105732452A - Method for extracting phaffia rhodozyma intracellular astaxanthin - Google Patents
Method for extracting phaffia rhodozyma intracellular astaxanthin Download PDFInfo
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- CN105732452A CN105732452A CN201610080243.XA CN201610080243A CN105732452A CN 105732452 A CN105732452 A CN 105732452A CN 201610080243 A CN201610080243 A CN 201610080243A CN 105732452 A CN105732452 A CN 105732452A
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- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C403/00—Derivatives of cyclohexane or of a cyclohexene or of cyclohexadiene, having a side-chain containing an acyclic unsaturated part of at least four carbon atoms, this part being directly attached to the cyclohexane or cyclohexene or cyclohexadiene rings, e.g. vitamin A, beta-carotene, beta-ionone
- C07C403/24—Derivatives of cyclohexane or of a cyclohexene or of cyclohexadiene, having a side-chain containing an acyclic unsaturated part of at least four carbon atoms, this part being directly attached to the cyclohexane or cyclohexene or cyclohexadiene rings, e.g. vitamin A, beta-carotene, beta-ionone having side-chains substituted by six-membered non-aromatic rings, e.g. beta-carotene
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Abstract
The invention discloses a method for extracting phaffia rhodozyma intracellular astaxanthin.Firstly, fermentation liquid of phaffia rhodozyma is concentrated to obtain bacterial sludge; a high-pressure homogeneous method is adopted to conduct thallus wall breaking, then, astaxanthin is extracted through a mixed solution of ethyl alcohol and acetic acid, reextraction is conducted through grease, a mixed phase is subjected to constant-temperature oscillation mixing, and after centrifugation layering, the grease phase on the upper layer is collected, and astaxanthin oil products rich in astaxanthin are obtained.The method is simple in technology, easy to operate and control and suitable for industrial production of astaxanthin.
Description
Technical field
The present invention relates to the technical field of astaxanthin, particularly relate to the technology of Astaxanthin extraction in red phaffia rhodozyma born of the same parents.
Background technology
Astaxanthin (Astaxanthin) is a kind of non-vitamin a source carotenoid, has extremely strong non-oxidizability and colorability, is widely used in fields such as food, medicine, feedstuff, cosmetics, health product.More of great interest because of the remarkable result in its prevention in many diseases (such as cardiovascular and cerebrovascular disease, parkinson's syndrome etc.) and treatment in recent years.
Astaxanthin can be divided into chemosynthesis astaxanthin and natural astaxanthin according to source.Chemical method synthesizing astaxanthin technical maturity, production cost is low, but the astaxanthin biological activity of synthesis is significantly lower, and there is certain residual toxicity, FDA(FDA) only ratify it and apply in feed industry, it is impossible to it is applied to medicine and field of food.Additionally, have begun to query the safety to aquaculture of the chemosynthesis astaxanthin in European Union, and draw up the application start discussion whether allowing the continuation of chemosynthesis astaxanthin in feedstuff.The production method of natural astaxanthin has two kinds: one to be extract from the Aquatic product leftover bits and pieces shell etc. of Eriocheir sinensis (shrimp), and the method exists the shortcomings such as raw material instability, Astaxanthin extraction rate is low, extraction process is complicated；Two is microorganism (Haematocoocus Pluvialls, phaffia rhodozyma etc.) fermenting and producing, content astaxanthin in haematococcus pluvialis cell is higher (up to dry cell weight 0.2% ~ 2.0%), but cultivation cycle length (up to more than 15 days), Biomass low (dry cell weight is usually no more than 3g/L), frustule extraction astaxanthin is also relatively difficult simultaneously, therefore, Haematococcus pluvialis production natural astaxanthin is inefficient.Than Haematocoocus Pluvialls, utilize that phaffia rhodozyma fermenting and producing astaxanthin has fast growth, fermentation period is short, fermentation technology is simple, High Density Cultivation can be carried out, need not the advantage such as illumination, be a kind of good natural astaxanthin source.
At present, on international market, the price of astaxanthin is about every kilogram of 2500-3000 dollar, and the whole world has more than the market capacity of 200,000,000 dollars every year, and presents ever-increasing trend.Therefore, widen the source of natural astaxanthin, study a kind of Astaxanthin extraction method of simple and effective, not only there is important researching value, more have wide practical use.
Summary of the invention
It is an object of the invention to provide a kind of Astaxanthin extraction method of simplicity, efficient, applicable industrialization promotion for red phaffia rhodozyma fermenting and producing astaxanthin.
In order to reach above-mentioned purpose, the solution of the present invention is:
The extracting method of astaxanthin in a kind of red phaffia rhodozyma born of the same parents, its step is as follows:
Step (1), fermentation liquid concentrates: the fermentation liquid of red phaffia rhodozyma, by concentrating, obtains bacterium mud；
Step (2), red phaffia rhodozyma thalline breaking cellular wall: thalline inactivates, regulate material liquid pH value, carry out high pressure homogenize breaking cellular wall；
Adopt sand mill, colloid mill, high pressure homogenizer etc. single or complex method is to red phaffia rhodozyma thalline breaking cellular wall；Physical method breaking cellular wall can avoid the organic solvent residual that organic solvent (such as dimethyl sulfoxide) breaking cellular wall produces, and simplifies the operation that organic solvent is removed, and saves production cost, increases economic efficiency；
Step (3), after thalline breaking cellular wall, carries out sand milling breakdown of emulsion；
Step (4), utilizes and extracts when dehydrated alcohol/acetic acid mixed liquor vacuum nitrogen, be centrifuged off bacterial chip, obtain astaxanthin crude extract；
Astaxanthin dissolubility in alcoholic solution is relatively low, and ethanol and acetic acid are mixed the dissolubility that can increase astaxanthin as Extraction solvent；Ethanol/acetic acid mixed liquor can change the permeability of phaffia rhodozyma cell, makes para chrome be discharged into outside born of the same parents, is dissolved in organic solvent system；Meanwhile, ethanol, acetic acid are harmless to people, animal, as the organic solvent extracting astaxanthin, can simplify the purification step of follow-up astaxanthin.Additionally, the boiling point of ethanol/acetic acid mixed liquor is relatively low, can be recycled by Distillation recovery, reduce industrial wastewater and generate, improve raw material availability；
Step (5), adds oils and fats, back extraction in astaxanthin crude extract, mixes through isothermal vibration, after centrifugal layering, collects upper strata oils and fats phase, obtains the oil extraction liquid of astaxanthin, and lower ethanol-Acetic Acid-Water is mutually by Distillation recovery ethanol and acetate component；
Astaxanthin is dissolvable in water the organic solvent such as ethanol, acetic acid, ethanol and acetic acid and water dissolves each other, and the ethanol-Acetic Acid-Water in system compares difficult removal, and oils and fats can help system layering, is formed biphase clearly；Meanwhile, astaxanthin is embedded in the stability that can keep astaxanthin in the middle of oils and fats better.
Above-mentioned strain is red phaffia rhodozymaPhaffiarhodozyma/XanthophyllomycesdendrorhousStrain.
In described step (2), bacterium mud is warming up to 68 ~ 75 DEG C of inactivation 20 ~ 40min.
In described step (2), after thalline inactivation, adding organic acid and regulate material liquid pH value to 4.0 ~ 6.0, the organic acid used is citric acid, malic acid, tartaric acid or oxalic acid.
In described step (2), high pressure homogenize breaking cellular wall is homogenizing 1 ~ 2 time under 80 ~ 100MPa pressure condition, it is to avoid the organic solvent residual that conventional organic solvents breaking cellular wall produces, and simplifies the purification step of Astaxanthin extraction.
In described step (4), the ethanol used/acetic acid mixed liquor is (v:v) ethanol by volume: acetic acid=1:1 ~ 3 mixing, and the ratio of Phaffia Rhodozyma weight (g) and ethanol/acetic acid mixed liquor volume (ml) is 1:10 ~ 15.
In described step (4), Extracting temperature 60 ~ 75 DEG C, extract 3 ~ 4h when vacuum nitrogen, extract 2～3 times, united extraction liquid is astaxanthin crude extract.
In described step (5), ethanol-Acetic Acid-Water in astaxanthin crude extract is removed by method mutually that utilize oils and fats back extraction, selected oils and fats is that DHA algal oil, high linoleic acid sunflower oil, soybean oil, Semen Lini oil be single or compound oil, and the volume ratio of oils and fats and astaxanthin crude extract is 1:0.2～1.0.
In described step (5), oils and fats back extraction is to make feed liquid mix homogeneously at 55 ~ 75 DEG C of isothermal vibration 30 ~ 60min.
After adopting such scheme, the present invention is concentrated by fermentation liquid, high pressure homogenize breaking cellular wall, ethanol/acetic acid mixed liquor slightly carries, the steps such as oils and fats back extraction, achieve and rapidly and efficiently extract the purpose of astaxanthin in red phaffia rhodozyma born of the same parents, and effectively keep the biological activity of astaxanthin, maintain stablizing of astaxanthin product quality.
Detailed description of the invention
Present invention is disclosed a kind of method extracting astaxanthin in red phaffia rhodozyma born of the same parents, comprise the following steps:
Step (1) fermentation liquid concentrates: is concentrated by the fermentation culture of red phaffia rhodozyma, obtains bacterium mud；
Step (2) thalline inactivates: bacterium mud is warming up to 72 DEG C of inactivation 30min, then adds Fructus Citri Limoniae acid for adjusting pH value to 5.5；
The red phaffia rhodozyma thalline breaking cellular wall of step (3): 90MPa high pressure homogenize makes thalline breaking cellular wall 2 times in high pressure homogenizer, then carries out sand milling breakdown of emulsion；
Step (4) ethanol/acetic acid mixed liquor slightly carries astaxanthin: extract astaxanthin with ethanol/acetic acid (v:v=1:1) mixed liquor by solid-liquid ratio 1:15 after thalline breaking cellular wall, described solid-liquid ratio is Phaffia Rhodozyma weight and the ratio of ethanol/acetic acid mixed liquor volume, i.e. g/ml, 75 DEG C are extracted 4h, extracting 3 times, united extraction liquid is astaxanthin crude extract；
Step (5) DHA algal oil back extraction: add the DHA algal oil of 5 times of volumes in astaxanthin crude extract, through 65 DEG C of isothermal vibration 30min mix homogeneously, it is divided into two-layer after centrifugal, collect upper strata oils and fats phase, obtaining the DHA algal oil extract of astaxanthin, lower ethanol-Acetic Acid-Water is through Distillation recovery ethanol and acetic acid.
The method technique of the present invention is simple, efficiently, it is easy to operation and control, it is adaptable to the industrialization of astaxanthin produces.
Claims (9)
1. the extracting method of astaxanthin in red phaffia rhodozyma born of the same parents, it is characterised in that step is as follows:
Step (1), the fermentation liquid of red phaffia rhodozyma, by concentrating, obtains bacterium mud；
Step (2), thalline inactivates, and regulates material liquid pH value, carries out high pressure homogenize breaking cellular wall；
Step (3), after thalline breaking cellular wall, carries out sand milling breakdown of emulsion；
Step (4), utilizes and extracts when dehydrated alcohol/acetic acid mixed liquor vacuum nitrogen, be centrifuged off bacterial chip, obtain astaxanthin crude extract；
Step (5), adds oils and fats, back extraction in astaxanthin crude extract, mixes through isothermal vibration, after centrifugal layering, collects upper strata oils and fats phase, obtains the oil extraction liquid of astaxanthin, and lower ethanol-Acetic Acid-Water is mutually by Distillation recovery ethanol and acetate component.
2. the extracting method of astaxanthin in a kind of red phaffia rhodozyma born of the same parents according to claim 1, it is characterised in that: above-mentioned strain is red phaffia rhodozymaPhaffiarhodozyma/XanthophyllomycesdendrorhousStrain.
3. the extracting method of astaxanthin in a kind of red phaffia rhodozyma born of the same parents according to claim 1, it is characterised in that in step (2), bacterium mud is warming up to 68 ~ 75 DEG C of inactivation 20 ~ 40min.
4. the extracting method of astaxanthin in a kind of red phaffia rhodozyma born of the same parents according to claim 1, it is characterized in that in step (2), after thalline inactivation, adding organic acid and regulate material liquid pH value to 4.0 ~ 6.0, the organic acid used is citric acid, malic acid, tartaric acid or oxalic acid.
5. the extracting method of astaxanthin in a kind of red phaffia rhodozyma born of the same parents according to claim 1, it is characterised in that in step (2), high pressure homogenize breaking cellular wall is homogenizing 1 ~ 2 time under 80 ~ 100MPa pressure condition.
6. the extracting method of astaxanthin in a kind of red phaffia rhodozyma born of the same parents according to claim 1, it is characterized in that in step (4), when astaxanthin slightly carries, ethanol/the acetum used is ethanol by volume: acetic acid=1:1 ~ 3 mixing, the ratio of Phaffia Rhodozyma weight and ethanol/acetic acid mixed liquor volume is 1:10 ~ 15.
7. the extracting method of astaxanthin in a kind of red phaffia rhodozyma born of the same parents according to claim 1, it is characterised in that in step (4), Extracting temperature 60 ~ 75 DEG C, extract 3 ~ 4h when vacuum nitrogen, extract 2～3 times, united extraction liquid is astaxanthin crude extract.
8. the extracting method of astaxanthin in a kind of red phaffia rhodozyma born of the same parents according to claim 1, it is characterized in that in step (5), ethanol-Acetic Acid-Water in astaxanthin crude extract is removed by method mutually that utilize oils and fats back extraction, selected oils and fats is that DHA algal oil, high linoleic acid sunflower oil, soybean oil, Semen Lini oil be single or compound oil, and the volume ratio of oils and fats and astaxanthin crude extract is 1:0.2～1.0.
9. the extracting method of astaxanthin in a kind of red phaffia rhodozyma born of the same parents according to claim 1, it is characterised in that in step (5), oils and fats back extraction is to make feed liquid mix homogeneously at 55 ~ 75 DEG C of isothermal vibration 30 ~ 60min.
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Cited By (7)
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| CN108813527A (en) * | 2018-04-11 | 2018-11-16 | 华南理工大学 | A kind of flavouring of astaxanthin-containing and preparation method thereof |
| CN108865891A (en) * | 2018-07-19 | 2018-11-23 | 威海利达生物科技有限公司 | A kind of method of phaffia rhodozyma cell mechanical breaking-wall method |
| CN110407659A (en) * | 2019-08-05 | 2019-11-05 | 嘉必优生物技术（武汉）股份有限公司 | A kind of Extraction of carotenoid pigment method |
| CN111466496A (en) * | 2020-03-10 | 2020-07-31 | 广东恒兴饲料实业股份有限公司 | Color-mixing compound feed capable of improving body color of African penaeus monodon and improving quality of African penaeus monodon |
| CN111793014A (en) * | 2020-07-27 | 2020-10-20 | 中国海洋大学 | Method for preparing water-soluble astaxanthin and astaxanthin aqueous solution prepared by method |
| CN114751848A (en) * | 2022-04-14 | 2022-07-15 | 厦门昶科生物工程有限公司 | Method for preparing astaxanthin essential oil based on phaffia rhodozyma |
| IT202200004247A1 (en) * | 2022-03-07 | 2023-09-07 | Univ Degli Studi Roma La Sapienza | METHOD OF EXTRACTION AND RECOVERY OF ASTAXANTHIN FROM BIOMASS |
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| CN108813527A (en) * | 2018-04-11 | 2018-11-16 | 华南理工大学 | A kind of flavouring of astaxanthin-containing and preparation method thereof |
| CN108865891A (en) * | 2018-07-19 | 2018-11-23 | 威海利达生物科技有限公司 | A kind of method of phaffia rhodozyma cell mechanical breaking-wall method |
| CN110407659A (en) * | 2019-08-05 | 2019-11-05 | 嘉必优生物技术（武汉）股份有限公司 | A kind of Extraction of carotenoid pigment method |
| CN110407659B (en) * | 2019-08-05 | 2023-04-07 | 嘉必优生物技术（武汉）股份有限公司 | Carotenoid extraction method |
| CN111466496A (en) * | 2020-03-10 | 2020-07-31 | 广东恒兴饲料实业股份有限公司 | Color-mixing compound feed capable of improving body color of African penaeus monodon and improving quality of African penaeus monodon |
| CN111793014A (en) * | 2020-07-27 | 2020-10-20 | 中国海洋大学 | Method for preparing water-soluble astaxanthin and astaxanthin aqueous solution prepared by method |
| CN111793014B (en) * | 2020-07-27 | 2021-07-13 | 中国海洋大学 | Method for preparing water-soluble astaxanthin and astaxanthin aqueous solution prepared by method |
| IT202200004247A1 (en) * | 2022-03-07 | 2023-09-07 | Univ Degli Studi Roma La Sapienza | METHOD OF EXTRACTION AND RECOVERY OF ASTAXANTHIN FROM BIOMASS |
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| CN114751848A (en) * | 2022-04-14 | 2022-07-15 | 厦门昶科生物工程有限公司 | Method for preparing astaxanthin essential oil based on phaffia rhodozyma |
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